Efficacy of a novel reverse transcriptase-polymerase chain reaction (RT-PCR) for detecting Toxoplasma gondii bradyzoite gene expression in human clinical specimens.
Efficacy of a novel reverse transcriptase-polymerase chain reaction (RT-PCR) for detecting Toxoplasma gondii bradyzoite gene expression in human clinical specimens.
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新型逆转录聚合酶链反应（RT-PCR）检测人类临床标本中弓形虫缓殖子基因表达的功效。
DOI：
10.1006/mcpr.2001.0394
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发表时间：
2002
影响因子：
3.3
通讯作者：
C. Contini
中科院分区：
文献类型：
--
作者：
R. Cultrera;S. Seraceni;C. Contini
应用逆转录-聚合酶链反应（RT-PCR）检测艾滋病（AIDS）弓形虫脑炎（TE）患者脑脊液（CSF）中弓形虫缓殖子和速殖子特异性基因的转录水平，以区分无症状潜伏感染和再激活感染。并与巢式DNA扩增（n）-PCR进行了比较。用RT-PCR方法检测了代表性T.从首次发病（11例）或复发（8例）的TE的AIDS患者获得的CSF中研究弓形虫囊性基质（MAG 1）或缓殖子特异性（SAG 4）基因。并对速殖子特异性基因SAG 1的mRNA表达进行了研究。新设计的寡核苷酸引物和探针用于RT-PCR和n-PCR检测，其识别开放阅读MAG 1序列内的212 bp片段。Oligo-dT引物的cDNA合成出现了一个合适的方法，为随后的分析，通过n-PCR。RT-PCR比n-PCR具有更高的敏感性和特异性。MAG 1和SAG 4基因分别在8例（100%）和6例（75%）TE复发患者中检测到表达，而SAG 1检测到7例（63%）TE首次发作患者。结果表明，RT-PCR方法能够鉴定T.尤其是在TE复发风险患者中。
A reverse transcriptase-polymerase chain reaction (RT-PCR) assay, was performed to evaluate the transcription degree of bradyzoite- or tachyzoite-specific genes of Toxoplasma gondii on cerebrospinal fluid (CSF) specimens from AIDS patients with toxoplasmic encephalitis (TE), and to distinguish an asymptomatic latent infection from a reactivated disease. This method was compared with nested DNA amplification (n)-PCR. The mRNA expression of the representative T. gondii cystic matrix (MAG1) or bradyzoite-specific (SAG4) genes was investigated on CSF obtained from AIDS patients with first episode (no. 11) or relapse (no. 8) of TE. The mRNA expression of tachyzoite-specific (SAG1) gene was also studied. New designed oligonucleotide primers and probes, which identify a 212 bp fragment inside to the open reading MAG1 sequence, were employed in both RT-PCR and n-PCR assays. Oligo-dT primed cDNA synthesis appeared a suitable method for subsequent analysis by n-PCR. RT-PCR has been shown to be more sensitive and specific than n-PCR. MAG1 and SAG4 gene expression was detected in 8 (100%) and 6 (75%) patients with TE relapses, respectively, while SAG1 detected 7 (63%) patients with TE first episode. These findings suggest that RT-PCR method is able to identify the bradyzoite stage of T. gondii especially in patients who are at risk for TE relapse.
克隆编码 28 千道尔顿弓形虫抗原的 cDNA。
影响因子：
1.5
作者：
Prince,JB;Araujo,FG;Remington,JS;Burg,JL;Boothroyd,JC;Sharma,SD
通讯作者：
Sharma,SD
影响因子：
3.5
作者：
Yang, SM;Parmley, SF
通讯作者：
Parmley, SF
影响因子：
4.4
作者：
J. L. Burg;D. Perelman;L. Kasper;P. Ware;J. Boothroyd
通讯作者：
J. L. Burg;D. Perelman;L. Kasper;P. Ware;J. Boothroyd